The prevalence of near-infrared (NIR) imaging within the multi-disciplinary field of molecular imaging has grown in recent years. This type of imaging has intrinsic advantages (e.g., low cost, non-invasive) when compared to other modalities such as positron emission tomography, magnetic resonance imaging, and computed tomography. However, several challenges remain before fluorescence NIR tomography algorithms combined with a mono-modal clinical device can provide quantitatively reliable information.